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RANK-Fc inhibits malignancy via inhibiting ERK activation and evoking caspase-3-mediated anoikis in human osteosarcoma cells

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Abstract

Osteosarcoma (OS) is the most common primary malignant bone tumour, mainly afflicting the young. While there has been substantial improvement in treatment of OS with surgery and chemotherapy in the past two decades, this disease remains a significant health problem, warranting efforts to find better therapeutic options. In this study, we examined the RANK/RANKL axis in OS cells, using a RANK-Fc protein to perturb this coupling in an effort to reduce OS cell growth. RANK-Fc suppressed OS cell migration (P < 0.005), invasion ability (P < 0.05), and anchorage-independent ability in collagen-1 gel (P < 0.005) following induction of anoikis and activation of caspase-3. OS cell proliferation was not perturbed by RANK-Fc. The anti-invasion and anti-metastasis capability of RANK-Fc is attributed to reduced extracellular signal-regulated protein kinase (ERK) signaling via RANK-Fc, though activation of NFκB, and altered expression of Akt, p38, JNK, and matrix metalloproteinase (MMP)-2 and -9 were ruled out. In vivo, activity of the RANK-Fc against OS cell migration and invasion was confirmed in a model strictly monitoring metastasis. Thus, RANK-Fc, given its ability to directly reduce OS aggression, is a potential drug candidate.

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References

  1. Clark JC, Dass CR, Choong PF (2008) A review of clinical and molecular prognostic factors in osteosarcoma. J Cancer Res Clin Oncol 134:281–297

    Article  CAS  PubMed  Google Scholar 

  2. Ta HT, Dass CR, Choong PF, Dunstan DE (2009) Osteosarcoma treatment: state of the art. Cancer Metastasis Rev 28:247–263

    Article  PubMed  Google Scholar 

  3. Mori K, Le Goff B, Berreur M et al (2007) Human osteosarcoma cells express functional receptor activator of nuclear factor-kappa B. J Pathol 211:555–562

    Article  CAS  PubMed  Google Scholar 

  4. Woods NT, Yamaguchi H, Lee FY, Bhalla KN, Wang HG (2007) Anoikis, initiated by Mcl-1 degradation and Bim induction, is deregulated during oncogenesis. Cancer Res 67:10744–10752

    Article  CAS  PubMed  Google Scholar 

  5. Jones DH, Nakashima T, Sanchez OH et al (2006) Regulation of cancer cell migration and bone metastasis by RANKL. Nature 440:692–696

    Article  CAS  PubMed  Google Scholar 

  6. Thomas RJ, Guise TA, Yin JJ et al (1999) Breast cancer cells interact with osteoblasts to support osteoclast formation. Endocrinology 140:4451–4458

    Article  CAS  PubMed  Google Scholar 

  7. Ignatoski KM, Escara-Wilke JF, Dai JL et al (2008) RANKL inhibition is an effective adjuvant for docetaxel in a prostate cancer bone metastases model. Prostate 68:820–829

    Article  CAS  PubMed  Google Scholar 

  8. Dass CR, Ek ET, Contreras KG, Choong PF (2006) A novel orthotopic murine model provides insights into cellular and molecular characteristics contributing to human osteosarcoma. Clin Exp Metastasis 23:367–380

    Article  PubMed  Google Scholar 

  9. Dass CR, Nadesapillai AP, Robin D et al (2005) Downregulation of uPAR confirms link in growth and metastasis of osteosarcoma. Clin Exp Metastasis 22:643–652

    Article  CAS  PubMed  Google Scholar 

  10. Ek ET, Dass CR, Contreras KG, Choong PF (2007) Inhibition of orthotopic osteosarcoma growth and metastasis by multitargeted antitumor activities of pigment epithelium-derived factor. Clin Exp Metastasis 24:93–106

    Article  CAS  PubMed  Google Scholar 

  11. Ek ET, Dass CR, Contreras KG, Choong PF (2007) PEDF-derived synthetic peptides exhibit antitumor activity in an orthotopic model of human osteosarcoma. J Orthop Res 25:1671–1680

    Article  CAS  PubMed  Google Scholar 

  12. Zhou H, Choong P, McCarthy R et al (1994) In situ hybridization to show sequential expression of osteoblast gene markers during bone formation in vivo. J Bone Miner Res 9:1489–1499

    Article  CAS  PubMed  Google Scholar 

  13. Dass CR, Khachigian LM, Choong PF (2008) c-Jun is critical for the progression of osteosarcoma: proof in an orthotopic spontaneously metastasizing model. Mol Cancer Res 6:1289–1292

    Article  CAS  PubMed  Google Scholar 

  14. Dass CR, Choong PF (2008) uPAR mediates anticancer activity of PEDF. Cancer Biol Ther 7:1262–1270

    Article  CAS  PubMed  Google Scholar 

  15. Dass CR, Choong PF (2007) Zoledronic acid inhibits osteosarcoma growth in an orthotopic model. Mol Cancer Ther 6:3263–3270

    Article  CAS  PubMed  Google Scholar 

  16. Fahmy RG, Dass CR, Sun LQ, Chesterman CN, Khachigian LM (2003) Transcription factor Egr-1 supports FGF-dependent angiogenesis during neovascularization and tumor growth. Nat Med 9:1026–1032

    Article  CAS  PubMed  Google Scholar 

  17. Ta HT, Dass CR, Larson I, Choong PF, Dunstan DE (2009) A chitosan hydrogel delivery system for osteosarcoma gene therapy with pigment epithelium-derived factor combined with chemotherapy. Biomaterials 30:4815–4823

    Article  CAS  PubMed  Google Scholar 

  18. Mehlen P, Puisieux A (2006) Metastasis: a question of life or death. Nat Rev Cancer 6:449–458

    Article  CAS  PubMed  Google Scholar 

  19. Mundy GR (2002) Metastasis to bone: causes, consequences and therapeutic opportunities. Nat Rev Cancer 2:584–593

    Article  CAS  PubMed  Google Scholar 

  20. Wittrant Y, Theoleyre S, Couillaud S et al (2003) Regulation of osteoclast protease expression by RANKL. Biochem Biophys Res Commun 310:774–778

    Article  CAS  PubMed  Google Scholar 

  21. Koistinen P, Pulli T, Uitto VJ et al (1999) Depletion of alphaV integrins from osteosarcoma cells by intracellular antibody expression induces bone differentiation marker genes and suppresses gelatinase (MMP-2) synthesis. Matrix Biol 18:239–251

    Article  CAS  PubMed  Google Scholar 

  22. Xin ZF, Kim YK, Jung ST (2009) Risedronate inhibits human osteosarcoma cell invasion. J Exp Clin Cancer Res 28:105

    Article  PubMed  CAS  Google Scholar 

  23. Frisch SM, Screaton RA (2001) Anoikis mechanisms. Curr Opin Cell Biol 13:555–562

    Article  CAS  PubMed  Google Scholar 

  24. Martin TJ, Mundy GR (2007) Bone metastasis: can osteoclasts be excluded? Nature 445:E19–E20

    Article  CAS  PubMed  Google Scholar 

  25. Hughes AE, Ralston SH, Marken J et al (2000) Mutations in TNFRSF11A, affecting the signal peptide of RANK, cause familial expansile osteolysis. Nat Genet 24:45–48

    Article  CAS  PubMed  Google Scholar 

  26. Sparks AB, Peterson SN, Bell C et al (2001) Mutation screening of the TNFRSF11A gene encoding receptor activator of NF kappa B (RANK) in familial and sporadic Paget’s disease of bone and osteosarcoma. Calcif Tissue Int 68:151–155

    Article  CAS  PubMed  Google Scholar 

  27. Bu R, Borysenko CW, Li Y et al (2003) Expression and function of TNF-family proteins and receptors in human osteoblasts. Bone 33:760–770

    Article  CAS  PubMed  Google Scholar 

  28. Tan ML, Choong PF, Dass CR (2009) Osteosarcoma: Conventional treatment vs. gene therapy. Cancer Biol Ther 8:106–117

    CAS  PubMed  Google Scholar 

  29. Lamoureux F, Richard P, Wittrant Y et al (2007) Therapeutic relevance of osteoprotegerin gene therapy in osteosarcoma: blockade of the vicious cycle between tumor cell proliferation and bone resorption. Cancer Res 67:7308–7318

    Article  CAS  PubMed  Google Scholar 

  30. Emery JG, McDonnell P, Burke MB et al (1998) Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL. J Biol Chem 273:14363–14367

    Article  CAS  PubMed  Google Scholar 

  31. Miller RE, Branstetter D, Armstrong A et al (2007) Receptor activator of NF-kappa B ligand inhibition suppresses bone resorption and hypercalcemia but does not affect host immune responses to influenza infection. J Immunol 179:266–274

    CAS  PubMed  Google Scholar 

  32. Lacey DL, Timms E, Tan HL et al (1998) Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93:165–176

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by funding from the Australian Orthopaedics Association, the Victorian Orthopaedics Research Trust, and St. Vincent’s Hospital Melbourne. The authors thank Mei Lin Tan for help with some of the immunoblots.

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Correspondence to Crispin R. Dass.

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Akiyama, T., Choong, P.F.M. & Dass, C.R. RANK-Fc inhibits malignancy via inhibiting ERK activation and evoking caspase-3-mediated anoikis in human osteosarcoma cells. Clin Exp Metastasis 27, 207–215 (2010). https://doi.org/10.1007/s10585-010-9319-y

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  • DOI: https://doi.org/10.1007/s10585-010-9319-y

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